Medical application for heparin and related molecules

ABSTRACT

New uses for heparin, or heparin-like compounds are described that encompass preserving and healing of cells and cell functions arising from transplantations, circumcisions, dermatitides, fissures, fistulas, stimulation of epithelial growth, keloid prevention, cold injuries, pathology and forensic diagnosis, myocardium, trauma, decubitus ulcers, psoriasis, poisonings, insect and snake bites, corrosive ingestions, the &#34;bends,&#34; space-travel sickness, brain and heart nerve conduction electrical dysrhythmias, pulmonary respiratory distress, blood and blood products, ulcerative colon lesions, interstitial cystitis, and related cosmetic uses. The uses are realized by applying the compounds either in solution, or in the form of a cream or aerosol, preferably at a pH of about 5.5, in an effective amount and for a time sufficient to effect treatment. Generally, the concentration of heparin or heparin-like compounds will be in the range of 1500 to 5000 international units per milliliter. Clinical assays are also described for determining the amount of heparin that should be used in those instances where the effective concentration is not known.

BACKGROUND OF THE INVENTION

Heparin is a polysaccharide present in a variety of organs and tissues,particularly liver, lung, and the large arteries. It is a polymer ofrepeating units of D-glucuronic acid usually having an O-sulfate groupat C-2 and D-glucosamine N-sulfate with an additional O-sulfate group atC-6. Both the linkages of the polymer are alternating -1,4, and it isthought that seven of the eight glucuronic acid residues in the polymerexhibit an O-sulfate group. The postulated repeating unit is shown asfollows; ##STR1## as defined by Jaques, Science, 206, 529 (1979) andRawn, Biochemistry, 297-298 (1983). Heparin is commonly provided as thesodium salt, sometimes referred to as sodium heparinate or heparinsodium, as indicated in U.S. Pat. No. 3,062,716 (Montaudraud) and as"Entry 4543" in The Merck Index, 672 (10th edn. 1983).

One of the major properties of heparin which has been recognized forsome time is its ability to prolong the clotting time of blood both invivo and in vitro. Less well known properties of heparin are that itstimulates healing of burn wounds and its ability to reverse ischemicmyocardial injuries. For instance, Saliba et al. in J. Amer. Med.Assoc., 225, 261 (1973) describe the application of heparin-administeredparenterally and topically to humans suffering from second and thirddegree burns. Heparin both relieves the pain and prevents the initialburn size from expanding. Most important, there is enhancedrevascularization, granulation and re-epithelialization. The effect ofheparin on burn wound healing is critically dose-related, dose-dependentand pH regulated (Saliba, Thrombosis and Haemostasis, 40, No. 1 (1978)).Most applications of heparin as a therapeutic agent have usedconcentrations in the range of 20,000 to 40,000 units/per milliliter forintravenous or subcutaneous modes of presentation, while for topicalapplications concentrations in the range of 5,000 to 10,000 units/permilliliter have been employed. The beneficial effect of heparin onreversing myocardial ischemic injury generally has been shown to requirethe administration of 10,000 to 100,000 units total of 5,000 to 10,000units/ml heparin, with favorable results being apparent acutely onelectrocardiograms and on cardiac enzymes monitored at 24 and 48 hours.

In addition to the above effects of heparin, the molecule is alsothought to exhibit antithrombin, antiplatelet-lysis, thrombolytic,antihistaminic, antiserotonin and antiproteolytic enzymatic activity.The mechanism whereby heparin exerts all these effects is not known.Lastly, heparin has also been shown to be effective in treating weepingpoison oak dermatitis (Saliba and Griner Aerospace Medicine, 41, 2, 179(1970)) and weeping ear eczema and acute tracheal bronchitis (Doughertyand Dolowitz, Amer. J. Cardiol. 14, 18 (1964)).

The following U.S. patents show various medical applications for heparinor related compounds. U.S. Pat. No. 3,062,716 shows a method of treatinghemorrhoids. U.S. Pat. No. 3,137,624 describes compositions for treatingdefective veins wherein one component is heparin. U.S. Pat. No.3,244,594 describes heparin compounds having antitumor activity. U.S.Pat. No. 3,151,025 describes a composition that rids the blood oflipoprotein molecules. U.S. Pat. No. 4,039,665 shows a method ofcomposition containing heparin for eradication of venous blemishes.Lastly, U.S. Pat. No. 4,390,532 shows another composition with heparinthat is useful for topical applications in treating wrinkles, acne andandrogen baldness.

SUMMARY OF THE INVENTION

It is an object of this invention to describe new uses for heparin andheparin related molecules, particularly heparin sulfate, heparin acidsand the like, that rely on the hithertofore unappreciated anticellulardestructive effects of these molecules. Over a well defined range ofheparin concentrations and pHs, particularly acidic pH's, it is shownthat there is an inhibition of cellular destruction which in turnfacilitates healing of wounded cells, tissues or organs.

This patent describes a method of using heparin and heparin relatedsubstances to facilitate transplantation in humans and animals bypreserving the transplant prior to removal from a donor and duringtransportation and surgical transplantation into the recipient. Heparinprevents cellular breakdown and destruction of the transplant, minimizesrejection and enhances physiological function of the transplant(s). Thisin turn obviates or reduces the need to use immunosuppressants, therebylessening morbidity and mortality.

By transplant it will be understood that reference is made to any humanor animal body part, cell, tissue or organ removed from a human oranimal donor and placed for use on, in or within a human or animalrecipient. It is a particular object of the invention to show thatbecause heparin and related compounds efficaciously prolong the in vitrolifetime of organs removed from transplant donors, it enables organs tobe transplanted into a recipient after the organ has been out of thedonor for a longer time than has been hithertofore been possible.

It is yet a further object of this invention to show that heparin, andrelated compounds, are efficacious in stimulating healing of skin woundsgenerally, but particularly including those arising from circumcision,cold injuries, dermatitides, fissures, fistulas, stimulation ofepithelial growth, keloid prevention, cold injuries, pathology andforensic diagnosis, myocardium, trauma, decubitus ulcers, psoriasis,poisonings, insect and snake bites, corrosive ingestions, the "bends,"space-travel sickness, brain and heart nerve conduction electricaldysrhythmias, pulmonary respiratory distress, blood and blood products,ulcerative colon lesions, interstitial cystitis and related cosmeticuses.

It will be understood by those skilled in the art that the scope ofapplications of heparin is not limited to the particular injuriesdescribed above. It is expected that it will be useful for simulatinghealing of virtually any tissue that requires re-epithelializationand/or revascularization when these processes are prevented or retardedfrom occurring due to the release of cellular chemicals exhibitingcytotoxic activity, and complicated by ischemia and infection.

BRIEF DESCRIPTION OF THE DRAWINGS

The Figures of the drawing are sets of photographs of the various stagesof the healing process for patients treated in accordance with thisinvention. Each of the sets is described in, respectively, Examples I,III, VII, VIII and X.

DETAILED DESCRIPTION OF THE INVENTION

Underlying the novel uses of heparin or chemically related moleculesdescribed herein is that they are capable, under suitable conditions ofpH and concentration, of retarding or preventing cellular-destructivebiochemical events which are associated both with slow wound healing andwith the rapid destruction of organs once removed from the body. Heparinalso stimulates endothelial cell migration and proliferation in ischemictissue. By preventing cellular and tissue destruction and by enhancingneoangiogenesis in ischemic areas it is possible to promote rapidrevascularization, granulation and re-epithelialization associated witha wound and shorten healing time and improve final functional andcosmetic results. Similarly it permits organ transplants using organsthat are have been removed from the donor for longer times thanhithertofore possible. These two types of medical problems are thepreterred, but not the only new uses that are beneficially treated withheparin.

Included within the category of skin wounds is ischemic traumatic woundswhich are beneficially treated with heparin to prevent vascular necrosisuntil an adequate vascular supply can be re-established. An example ofthis type of skin wound would be a traumatic amputation.

Other ischemic applications include myocardial ischemia or injury at anystage and arrhythmias. Treatment can consist of venous infusion using aperipheral vein or the administration of heparin by an in-dwellingcatheter at the base of the aorta or in coronary artery(ies) directly.The use of heparin and heparin related substances preserves ischemicmyocardium which in turn preserves heart function. Heparin selectivelyprevents and corrects cardiac arrhythmias. Although heparin does notlyse blood clots as effectively as streptokinase and urokinase, it hasmultiple anticellular destructive effects not found in those chemicalsthat preserve and heal ischemic myocardium.

Another application of heparin is to treat circumcision wounds. Theideal concentration is 2,500 International Units/Milliliter (I.U./ml).While a variety of treatment modalities will perform satisfactorily, thepreferred treatment consists of applying 0.5 ml of heparin solutiondirectly on the penis after surgery and then applying a nonadheringdressing soaked with 1.0 ml of heparin solution to enhance healing,especially the rapid re-epithelialization of the surgically denudedglans penis and residual foreskin surfaces, and to prevent adherence andadhesion of the denuded non-healed surfaces. Currently the adheringforeskin in many boys must be repeatedly and painfully peeled off itsattachment to the glans intermittently, for months or yearspost-circumcision, before there is proper healing. The heparin treatmentcan be used whenever the adhering foreskin is separated from itsattachment to the glans penis surface, days, months or years aftercircumcision. A treatment package would contain an individual 3 ml vialof 2,500 I.U./ml sterile acidic heparin solution, a vial of sterilephysiological saline water, appropriate-size nonadherent pads and a meshcircular gauze to be used by surgeons and to be provided to a patient'sparents for post-circumcision care of the circumcised penis.

Another procedure that facilitates the re-epithelialization ofcircumcision wounds consists of topically applying heparin fromintestinal mucosa in solution at about 2,500 I.U./ml immediatelypostcircumcision. 0.5 ml of the solution is applied on the denudedsurfaces and 1.0 ml on the nonadhering bandage and 1.0 ml with each, ifany, changing of the bandage into final healing. The heparin soakednonadhering bandage dressing will be wrapped around the denuded surfacesand held in place with the circular gauze mesh dressing. Sterile waterwill be used to soak the old bandage prior to its removal from thehealing surface to prevent disruption of the healing new epithelialsurface.

Heparin protection of cellular destruction and enhancement ofrevascularization and heparin induced epithelialization andre-epithelialization can be favorably applied to treat dermatitides,fissures and fistulas that are difficult to heal or are refractory tocurrent methods of treatment. These can be healed by the addition ofheparin using the methods described for treating circumcisions whereinheparin is generally applied topically twice a day, and sometimessubcutaneously for a limited time in a limited dose. Further, heparincan be used to stimulate and facilitate in vivo re-epithelialization ofdiseased or injured or denuded body surfaces and in vitro for skinhomografts and heterografts, as well as to inhibit or prevent theformation of keloids.

Further uses of heparin and related molecules include skin injuries dueto cold temperature exposure, such as frostbite. The latter can betreated in any stage of the injury, especially prior to or during thewarm-upstage, as well as in the intermediate post thawing stage and forseveral hours or days thereafter as needed to effect complete recovery.The use of heparin in hypothermia as a preventive or as a treatment willreduce morbidity and mortality. This use will be invaluable to humansand animals exposed to environmental conditions where hypothermia can belife-threatening, including but not limited to mountain climbers,deep-water divers and individuals living in cold environments on earth.A self-administered subcutaneous injection of heparin, in 10,000-40,000I.U. doses, once or twice daily, would prevent bloodclotting andcellular-destruction and preserve and maintain life. A continuousintravenous infusion could be used with the dose monitored by healthpersonnel during medical treatment. For the final phases of recovery athome, the self-administered subcutaneous injections of 5,000 to 20,000I.U. daily would complete the treatment.

Sunburns or thermal burns are yet another class of skin treatments thatare efficaciously treated by heparin. Again, it will be appreciated thatwhile heparin is the preferred chemical, molecules having similaractivity will perform equally well.

In ongoing experiments, psoriasis has responded to treatment withheparin applied topically and parenterally and specifically is intendedto come within the scope of this application.

Heparin can be beneficially utilized in the area of organ transplants asits anticellular destructive activity essentially preserves organs thathave been removed from a donor for longer periods of time than hashithertofore been possible. This allows for organs, tissues or cells tobe removed and then flown to virtually any region of the earth for useby a needy recipient.

An exemplary procedure for preparing an organ for transplantation mayconsist of first infusing the intract donor with a bolus intravenousinjection of about 10,000 I.U. of heparin given just prior to initialsurgery. Preferably the solution will be buffered at a pH of about 5.5.Heparin anticoagulates the donor and neutralizes cellular-destructiveinflammation-producing biochemicals such as histamine, serotinin, etc.Next, surgically expose (the donor part(s) and the transplants andisolate the transplant's major arteries and veins, whenever possible.Use a soft-covered surgical clamp to occlude the transplant's principalartery at its most proximal origin and the greatest distance away fromits entrance into the donor part, the transplant. Using a sufficientquantity of about 1,000 I.U./ml intestinal source heparin solution ordextran solution infused through an eighteen gauze needle inserted intothe artery just beyond the clamp, flush out the blood in that artery andthe transplant and the exiting veins with the heparin or dextransolution until no blood remains. This will prevent intratransplantclotting of blood and the pathological sequellae of micro- andmacro-infarctions, and also remove donor antigens within thetransplant's blood vessels that later could sensitize the recipient.Then through the same needle in the major artery, fill the transplant'sblood vessels with heparin solution in about 20,000 I.U./mlconcentration, while progressively and quickly clamping at the mostdistant location, the exiting veins, retaining the 20,000 I.U./mlsolution within the artery, the vascular-space of the transplant and theexiting veins. Finally, the transplant can be removed from the donor,and put into a plastic sterile bag containing sterile heparin solution,preferably buffered at a pH of about 5.5 and containing about 5,000I.U./ml concentration, which has been warmed to a constant temperatureof 95° F. (35° C.) regulated automatically by theelectrically-warmed-transporting-blanket, for rapid transport to arecipient.

During surgery the recipient and the transplant are maintained in anacidosis which prevents breakdown of the transplant, and the recipientis heparinized at the conclusion of the surgery by continuousintravenous infusion to prevent initial destruction of the transplantedtransplant. After intravenous infusions have been terminated, thepatient is maintained in a heparinized condition post-operatively bysubcutaneous injections of heparin once or twice daily with about 5,000to about 20,000 I.U. doses, keeping the laboratory coagulation factorswithin therapeutic range. This can be carried out in a hospital byhealth professionals, or at home by the patient or relatives for anextended period of time to minimize organ rejection and enhancephysiological function of the transplant(s).

It will be appreciated by those skilled in the art that while presenttransplant techniques preserve organs at low temperatures, particularly4° C., that this may not be the optimal storage temperature. Therationale for keeping the organ at low temperature is to inhibitcellular degradative processes. Thus, it will be appreciated that anadditional appealing feature of the instant invention is that it ispossible to maintain the organ at physiological temperatures, or about37° C., because at this temperature the anti-cellular destructiveeffects of heparin are apparent. Consequently the physician is given achoice of preserving organs at low temperatures, or if this is notdesirable, the organ can be maintained at near physiologicaltemperatures.

Related to the preservation of organs is the short term in vitropreservation of tissues for pathological examination, or forensic orother uses. Particularly applicable may be the preservation of tissuesfor microscopic examination to aid in the evaluation of a particulardisease state. Thus, tissues can be removed in a biopsy procedure,treated with heparin without use of chemicals that kill the cells, andthen examined microscopically without fear that the examination willreflect the deterioration due to cellular destructive enzymes whichcould lead to a misdiagnosis. For example, a variety of tumors haveassociated membrane receptors that are susceptible to degradation oncethey are removed from the body. Such receptors are often foundassociated with breasts or ovarian tumors. Using conventionalpreservation techniques these receptors are often not detectable,consequently the histochemical nature of the patient's cancer is notdiagnosed, often with disastrous consequences. Without wishing to berestricted to a particular mechanism, it is thought that heparin orrelated chemicals facilitate forensic diagnosis by preventing thebreakdown and protecting and preserving cells, tissues, etc. for alimited time, so that a more accurate initial determination of thecondition or state of the cells, tissue, etc. can be ascertained ongross and microscopic examination or by physical or biochemicaldetermination.

Utilization of heparin to preserve tissues that are to be examinedmicroscopically in medical diagnosis can be achieved by incorporatingheparin in an amount of 1,500-5,000 international units per milliliterin a suitable physiologically balanced salt solution and contacting thesalt solution onto the tissue, or in other ways immersing the tissue inthe solution. This procedure may be used alone or with currently usedchemical fixation methods. However, it is probably most suitably used onfrozen sectioned biopsy material which is susceptible to cellulardegradation in that the resolution afforded by the frozen sectiontechnique is negated if the tissue is first subjected to chemicalfixation. It will be further appreciated by those skilled in the artthat preservation of tissues will also find considerable application inthe forensic field.

I will now describe the preferred conditions under which heparin isbeneficially employed. The conditions most suited to the two generaltypes of medical applications will be described separately. The firstwill consist of conditions suited to facilitating wound healing,regardless of whether the wound is on the exterior or interior of anepithelial body surface. The second focuses on the conditions that mosteffectively retard organ, tissue, or cellular degradation once these areremoved from the body.

The first category of treatments, that is, interior or exteriorepithelial skin wounds, involves applying to the area of the wound about5000 international units per milliliter of heparin. This concentrationis most suited for topical applications wherein heparin is convenientlyadministered in the form of a solution or a cream or salve. A variety ofsuch creams or salves are well known to those skilled in the art. Ifheparin is desired to be administered throughout the body to aidinternal wound healing, this can be achieved by injecting heparinsolution at a concentration of about 10,000-40,000 international unitsper milliliter and maintaining this dose for several days or until suchtime as a beneficial result is apparent. I have found that for topicalapplications of heparin, it is most efficacious to apply the solution ina carrier having an acidic pH and particularly a pH of about 5.5. Forreasons that are presently unknown, the medically beneficial uses ofheparin are most apparent at acid pH's. Without wishing to be restrictedto a particular mode of action of heparin as applicable to the instantinvention, it is likely that this pH is favored because heparininteracts with and inactivates molecules involved in an inflammatoryreaction, particularly histamine and serotonin and proteolytic enzymesmost effectively at acid pH's. Since histamine is known to effectcellular destruction, its inhibition likely facilitates wound healing.

Further medical applications of heparin, hithertofore not known in theart, are similarly premised on the anticellular destructive activity ofthe molecule. Thus, for example, it is expected that heparin willpreserve blood and blood products thereby giving them a longer shelflife than is presently observed. Similarly, cell and tissue injuries dueto snake or insect bites, or ingestion of corrosive chemicals will alsobenefit from heparin application. In these instances, it is anticipatedthat heparin concentrations of about 1,500-5,000 international units permilliliter will be most efficacious.

As alluded to above, a variety of pharmaceutical carriers can beemployed with heparin to realize its beneficial uses. Such can be inliquid, solid or vapor form, and can be applied by injection, dripping,spreading or aerosol. A variety of solid pharmaceutical carriers can beimagined including starch, sugars, talc, manitol and the like. Again, itshould be stressed that such compositions preferably have an acidic pHat the site of injury, and particularly a pH of about 5.5. It will beunderstood that the term carrier is meant to include, in addition to theabove, buffered and non-buffered liquid.

It is important to note that for a particular medical application thatthe precise concentration of heparin that is most beneficial may bedetermined empirically. That is while concentration ranges are providedherein that are believed to be those that are most useful for aparticular application, in many instances these concentrations may notbe optimal, and that it may be desirous to have a procedure that can beutilized to determine the optimal heparin concentration.

It will be appreciated by those skilled in the art that there are atleast two assays that can be utilized to determine the optimal heparinconcentration. The first can be done in a clinical setting and relies onascertaining the immediate physiological changes caused by heparin.These are relief from pain, inflammatory erythema blanching andcessation of wound weeping. Cessation of pain and inflammatory erythemablanching occur within minutes of heparin application, and thus gives aready indication of whether a suitable concentration of heparin is beingutilized. Cessation of wound weeping occurs approximately 6 to 24 hoursafter treatment and serves as a further indication that the properamount of heparin is being utilized. In addition, other indicators thatan appropriate amount of heparin is being applied is that the wound siteundergoes revascularization and regranulation. These events occur laterin time than the prior events, and thus act as a correlative check toinsure that the proper amount of heparin is being employed. It will beapparent that based on both subjective doctor-patient communications andobjective observations by the physician that an empirically userulconcentration of heparin can be determined for a particular application.

It will be appreciated that while the minimally effective dose ofheparin can be determined by the above methods, that considerationshould also be given to insure that an excessive amount of heparin isnot administered. An excessive amount of heparin is that which no longerhas medically beneficial uses, and has significant anti-blood clottingactivity. The latter is readily ascertainable by monitoring a patient'sclot time. Generally clot times of 11/2-2 times longer than normalindicate that excessive doses of heparin are being used. Thus, bymonitoring the above parameters of pain erythema blanching, etc., andclotting time, the lower and upper limits of heparin concentration canbe ascertained.

A second procedure is based on determining the amount of histaminepresent at a particular wound site or that is associated with thecellular destructive processes ongoing in organs or tissues that havebeen removed from a donor prior to transplantation or microscopicexamination respectively. The procedure involves measuring the amount ofheparin needed to inactivate histamine present as a result of cellularinjury and consist of extracting histamine from the subject tissues ororgans and titrating it against heparin. Such procedures are well knownto those skilled in the art. Also the reaction conditions for titratinga particular amount of heparin needed to inactive histamineconcentrations in biological fluids and tissues are also well known.Particularly it is known that at a pH of about 5.5, one milligram ofhistamine binds to approximately 400 units of heparin. This relationshipenables a determination of approximately how much histamine is presentin the subject tissues or organs and consequently how much heparinshould be used.

The invention is described below with reference to particular examples.However, it will be understood by those skilled in the art that theinvention is not limited to either the materials or methods described inthese examples. It will be particularly apparent that the applicationsof heparin and related molecules are not restricted to humans but can beapplied to treating animals suffering from similar bodily disorders.Moreover, it will be readily apparent that there are numeroussubstitutions as to both the materials and methods that can be easilyimagined and that will substitute equally well for the materials andmethods described.

It will be further appreciated by those skilled in the art that whilethe mechanism by which heparin exerts its myriad effects is not known,it is nevertheless apparent that it maintains, promotes and protectsvital cellular functions. Consequently it is to be anticipated that inaddition to the uses described below, heparin will be found to haveother related uses. Moreover, as shown in some of the examples, heparinis also capable of enhancing as well as preserving and restoringcellular functions. Thus it is apparent that heparin is a key medicamentwith a truly wide range of medical uses and that it is the intent ofthis patent that applications of heparin based on its above-describedproperties are to be considered within the scope of this patent.

EXAMPLE I Caesarean Incision Cellulitis After Abscessed Dehiscence

A 30 year old female displaying an abscess-pocketed cellulitis resultingfrom complications of a Caesarean section and that extended across mostof the incision was treated with heparin. Prior to treatment the lesionwas infected, ischemic, necrotic and widely patent and it had a purulentpungent odor. FIG. 1A. The lesion was 13.5 cm long and 2.5 cm in depth(average). Heparin was administered subcutaneously in a 20,000 I.U.dose. After some purulent semi-liquid material was gently wiped out,40,000 I.U. of heparin was sprayed onto the necrotic infected walls.Then, after a thin plain sterile gauze pack was inserted, an additional60,000 1.U. of heparin was slowly applied to the gauze within thelesion. A single layer of Telfa pad dressing wa applied and taped to theabdominal wall at only three spots along the superior margin of theTelfa pad. Oral Ultracef was started in a 500 mg twice daily dose.Heparin treatment relieved the pain and it did not recur.

Six hours later, there were faint traces of blood on the gauze packing,and the surfaces of the lesion had the erythematous rubor of earlyrevascularization. FIG. 1B. The purulent exudate observed six hoursearlier had not returned and the previous pungent odor was not present.FIG. 1B. Slowly and repeatedly, over a twenty-minute period, the 5,000I.U./ml heparin solution was applied topically, 20,000 I.U. prior to,and 40,000 I.U. after, reinserting a new thin gauze packing.

The next day, at 24 hours after the onset of heparin treatment, theTelfa bandage was stained with dried clear secretions and reddish blood.The gauze packing was blood-ringed. FIG. 1C. The walls of the lesionwere more erythematous and revascularized, showed areas of earlygranulation between areas of whitish fibrous tissue. FIG. 1C. Overallthere was much less infection and no pungent odor. The patient had nopain. Then heparin solution was administered, 15,000 I.U. subcutaneouslyand 75,000 I.U. topically (40,000 I.U. prior to and 35,000 I.U. afterreinsertion of the thin gauze packing). The thin Telfa dressing waschanged twice daily, by physician or, at home, by her husband.

Starting day 2, at home, the patient's husband added topical heparinsolution to the dehisced incision and the gauze two or more times a day,including when he changed the gauze once or twice a day. The total dailyheparin dose applied by the husband was 25,000 I.U. for 3 days, then20,000 I.U. for 15 days, then 10,000 I.U. for 2 days. In the office anadditional 205,000 I.U. of heparin solution was applied topically intofinal healing, or approximately 20,000 I.U. daily on those days when itwas added. Thus the total heparin, administered topically by thephysician and by the husband, day 1 through day 22, was 732,000 I.U.Some topically-applied heparin promptly drained out before,theoretically, it could be efficiently utilized. With sloweradministration, a lesser amount of topical heparin might be aseffective.

Heparin was administered subcutaneously once a day by the physician inthe office or by the husband at home. The subcutaneous dose was 20,000I.U. daily, treatment days 1, 3, 5-16; and 10,000 I.U. daily, treatmentdays 17-22. The dose on day 2 was 15,000 I.U. The patient's husbandadministered the injections of heparin at home on treatment days 8, 9,12-16, 18-21, and 22. The total parenteral heparin dose days 1-22 was365,000 I.U.

On day 11, 22 hours after 20,000 I.U. of heparin was administeredsubcutaneously and 40,000 I.U. had been applied topically, theCoagulation Panel values were normal: Bleeding Time 4 minutes;Prothrombin Time 12, Control 12; ACTIVATED Partial Thromboplastin Time27, Control 28; and Platelet Count 227,000. Coagulation was not alteredby these doses of heparin.

The dehisced incision progressively revascularized, granulated in,closed and healed by re-epithelialization along the incision skinsurface, without complications, on the 29th day of treatment. Photograph2D shows the incision on treatment day 8.

The dehisced incision was widely patent and measured 13.5 cm in lengthand 2.5 cm in depth (average) when first observed. Treatment day 5, thedepth was 2.4 cm. The length was 11.25 cm and the depth was 1.8 ontreatment day 7. On treatment day 10, the length was 10.0 cm and thedepth was 1.1 cm. By the 17th day of treatment the length was 9.0 cm andthe depth was 0.7 cm. On day 23 the depth was 0.2 cm. The lesion wasclosed day 29.

Thus, an ischemic necrotic infected dehisced Caesarean-sectiontransverse suprapubic incision in a 30 year old woman that had notresponded to standard conventional treatment was healed by three weeksof daily topical heparin applied several times a day and once dailysubcutaneous heparin injections in 20,000 I.U. or 10,000 I.U. doses andoral cephalosporins. No other medical or surgical treatment wasnecessary.

EXAMPLE II Necrotic Infected Weeping Wound of Forearm

A 54 year old woman injured her left forearm in a water-skiing accident.The 5.1 by 5.6 cm wound, which was partially an abrasion and partiallyan avulsion of the skin on the dorsal surface, became infected. Onephysician she consulted had advised her to keep it moist withapplications of nitrofuradantin ointment. Three days later, a secondphysician had prescribed oral Penicillin VK and an unknown topicalointment to dry the weeping infected lesion. Neither treatment had beeneffective.

The 5.1 by 5.6 cm necrotic, purulent, avascular surface lesionsurrounded by an area of inflammatory erythema on the left forearm wassubsequently treated with heparin as follows. First, 10,000 I.U. ofheparin was injected into subcutaneous abdominal fat. Topical heparin, 4ml of 5,000 I.U./ml concentration, was slowly dripped onto the purulentnecrotic wound over a ten-minute period of time. After waiting another10 minutes to allow full exposure of the heparin to the wound, the areawas loosely bandaged with a dry nonadhering sterile Telfa pad. Anadditional quantity of heparin was sprayed on and soaked through theTelfa pad directly over the wound. The patient was given dicloxacillin250 mg, four times daily. Seven hours later, the Telfa pad was removedand, with it, a quantity of purulent material. The same treatment withtopical heparin was repeated.

The next morning, 24 hours after heparin treatment was started, thewound was clinically 90% free of infection. There was increased bloodflow within the wound, and there was no surrounding inflammatoryerythema. There was evidence of early granulation. Heparin subcutaneousdose was 10,000 I.U. and heparin topical dose was 2 ml of the 5,000I.U./ml solution. Telfa pad dressing and oral dicloxacillin werecontinued. The next day, 48 hours after initiating treatment, the woundwas estimated to be 50% well and rapid re-epithelialization was inprogress. Heparin was used topically, in 5,000 I.U. doses, for the lasttime. Twelve days after heparin treatment was initiated, there was newpink-colored skin over the entire area. No contractures developed. Thenew skin was only partially pigmented, but it was cosmetically pleasant.

EXAMPLE III Postoperative Cellulitis with Abscess-Sinus Tract

A 66 year old male displaying cellulitis involving the posterior lowerthird of his right leg, and having within the area of the cellulitis anabscess sinus tract was treated with heparin. The opening of the tractwas 20 mm above the calcaneus insertion of the achilles tendon. Theabscess sinus tract extended up into the leg along the achilles tendon,of which a thinned-down segment was visible within the ostia and sinus.The ostia was 30 mm in vertical diameter and the tract was 97 mm inlength. The greyish walls of the tract and ostia surface edges wereavascular and necrotic and covered with a purulent and pungent weepingfluid exudate. The purulent and pungent fluid was draining persistentlyfrom the ostia. FIG. 2A. It is important to note that this patient hadbeen treated for five months prior to heparin treatment using surgicaldebridement, oral and topical antibiotics and twice daily soaks withouteffect.

Heparin treatment was started midmorning, with 20,000 I.U. heparininjected into subcutaneous abdominal wall fat and 4 ml of the 5,000I.U./ml heparin solution (a total 20,000 I.U.) was slowly and repeatedlyapplied topically so that the heparin was placed within the abscesstract to rinse the wound; the heparin was dripped on the ostia opening;and finally, the heparin was used to saturate a sterile gauze packingtape that was inserted within the draining abscess tract. Oralantibiotics, dicloxacillin 500 mg, twice daily, and Geocillin 382 mg,three times a day, were continued by mouth. A sterile dry nonadheringTelfa pad was applied. Soaking of the leg in water or in antimicrobialwater solution was discontinued.

Eight hours later, there was an increased vascular-type rubor color tothe skin of the upper areas of the cellulitis, and less swelling of theankle and foot. There were faint traces of blood on the tape and at theostia and less drainage from the sinus tract. Treatment consisted of thetopical administration of heparin, including reinsertion of the heparinsolution-soaked packing tape within the sinus tract.

The next morning, 22 hours after the initial heparin treatment, theTelfa pad dressing and the gauze-tape sinus packing were soaked with aserous, pinkish, blood-tinged fluid. FIG. 2B. The drainage from theabscess sinus tract was less and it was nonpungent and less purulent innature. There was less tissue swelling and more rubor color to the skinof the ankle and heel.

Heparin was administered: 20,000 I.U. subcutaneously and a total 20,000I.U. topically within, and on, the abscess sinus tract and onto the tapeplaced within the tract. The tape was less deeply replaced.Subsequently, as the sinus tract filled in, the gauze tape packing wasreplaced progressively less deeply. At 32 hours, 10,000 I.U. of heparinwas topically applied.

The following morning, 46 hours after initiating heparin therapy, theswelling had subsided to the extent that the medial malleolus of thetibia was then prominent. Normal skin rubor had returned to the ankle,signalling a more normal blood flow. Traces of bright red blood were onthe packing that had been within the tract. Overall, there wasessentially minimal drainage, and it was not pungent or purulent.Subcutaneous heparin injection was 10,000 I.U., and 5,000 I.U. ofheparin was applied topically and repeated again at 56 hours.

The next morning, 72 hours after heparin treatment was started, theswelling was reduced by 40%. Portions of the tape had dark red-coloredblood. The length of the abscess tract was reduced from 97 mm to 50 mm.There was very little drainage. The skin blood supply on both sides ofthe ankle was vastly improved, as evidenced by the skin rubor and theprominent veins. The same doses of parenteral and topical heparin wereagain applied. Cultures of the sinus tract were taken and, after 48hours, were reported to be negative for bacterial or fungal growth.

Treatment with parenteral heparin, 10,000 I.U., was given the next 4days, then discontinued for 8 days. Topical heparin was continued, withdaily reduction in dose, into final healing.

At 118 hours, the reduced length of the packing tape is shown in FIG.2C, and the 60% reduction in swelling in FIG. 2D. The sinus tract wasprogressively filling in, and the ostia was circumferentially beingreduced in size by vascular granulation tissue. FIG. 2D.

The packing tape was less deeply replaced, and was no longer requiredafter the sixth day, as the tract had granulated in, and was then only ashallow depression within the ostia. The ostia also was rapidly reducingin size. But, for unknown reasons, healing of the ostia opening requiredmore days than the filling in of the cellulitis-abscess-sinus tract.Surface closure healing with heparin apparently awaited complete deephealing.

Fifteen days after treatment was started, when the ostia was almost 90%closed, the patient was seen by a plastic surgeon in the latter'soffice, for an appointment made prior to the starting of heparintreatment. Unaware of the heparin therapy, the plastic surgeon, withoutwarning, surgically debrided the lesion widely in the area of the nearlyhealed ostia. Blood promptly flooded from the freshly denuded area,attesting to the abundant revascularization that had taken place. Theplastic surgeon's intent was to remove surface epithelium so that itwould not cover over, before deep healing had been completed. Withoutprior knowledge of heparin's effects, the plastic surgeon had notrecognized that the sinus tract had already healed fully and that nosurface debridement was necessary at that point. Because he had expecteda relatively avascular response, he was surprised and apologetic. FIG.2F shows the ostia area 4 days after the office surgery. Unhappily, theskin surface healing was disrupted and delayed. Complete healing of thelesion that was expected in less than a month, required several weeksmore to re-epithelialize and become full-thickness skin. FIG. 2G showsthe lesion a month later.

Following this surgical debridement, parenteral heparin was startedagain, with 10,000 I.U. administered subcutaneously daily for 8 days.Topical heparin was administered in diminishing amounts until fullre-epithelialization was evident. The healing progressed slowly andsmoothly into final healing without further complications. Fullfunctional use of the leg and foot were possible when the wound washealed, and the cosmetic result was pleasant in appearance. FIG. 2Hshows the leg three years later.

EXAMPLE IV Cervical Cellulitis with Hypopharynx to Skin Fistula

A 55 year old male suffering from complications due to a laryngectomywas treated with heparin. The complications consisted of a cellulitisand a fistulous tract from his hypopharynx extending out the right sideof his neck. The cellulitis did not respond to treatment withantibiotics, and the fistulous tract did not respond to cauterizationwith silver nitrate, which was performed prior to heparin treatment by asurgeon five times. With every deglutition, masticated food and oralfluids exited via the fistula. The fistulous tract progressivelyenlarged.

Topical treatment with heparin was initiated by a physician, andcontinued after the first day by a registered nurse. Every 6 hours,topical heparin solution was placed within the fistulous tract until itwas full. Liquid erythromycin in a 250 mg per 5 ml dose was given via anasogastric tube every 6 hours. A dry sterile Telfa pad was applied overthe fistula ostia. No gauze packing was placed within the fistuloustract. No additional medical treatment was given and there was nofurther surgical treatment.

The fistulous drainage became progressively less and the previouspurulent pungent discharge was replaced by a scant clear fluid, whichwas occasionally tinged with a trace of blood. The swelling and doughyconsistency of the neck were progressively reduced and replaced bynormal pink firm tissue into the final healing phase. The greyishnecrotic surfaces of the fistula revascularized, granulated and theostia re-epithelialized progressively into final healing. The fistuloustract healed from the inside out, and only in the final phase of healingdid it close on the surface. After 11 days of topical heparin use, andthe day heparin treatment was stopped, the external ostia wasre-epithelialized with a thin layer of cells. Thereafter, the epitheliumprogressively thickened into normal thickness skin. No surfacecontractures developed. No local or systemic bleeding occurred. No painmedication was required.

EXAMPLE V Pyoderma-Cellulitis in a Pretibial Laceration

A 69 year old female presenting torn skin of the left leg above thetibia resulting from injury to a subcutaneous hematoma was treated withheparin. Heparin solution was applied twice daily to the infected,weeping, necrotic wound, by the physician in the office or by thepatient at home, for 12 days. Day 1, topical heparin dose was 20,000I.U., twice daily; reduced progressively to 2,500 I.U., twice daily, onday 12. Heparin, in 10,000 I.U. dose, was parenterally administered byinjection into subcutaneous fat, once daily for 3 days.

The wound revascularized, granulated, re-epithelialized and healedslowly. Antibiotics were discontinued after 18 days. A scab formed overthe wound. The scab fell off the fully healed area four weeks afterheparin therapy was initiated. No skin contractures developed and thecosmetic result was pleasant.

EXAMPLE VI Motorcycle Accident "Asphalt Burns" Arm, Leg, and Body

A 19 year old male having extensive asphalt "burns," abrasions andirregular skin avulsions of his right upper and lower extremity, flank,and hip resulting from a motorcycle accident was treated with heparin.Topical heparin solution, 8 ml of 5,000 I.U./ml concentration, wassprayed onto the "burns" twice daily, for 3 days; and 10,000 I.U. wassubcutaneously injected into fat, once daily, for 3 days. Erythromycinwas orally administered in 250 mg dose, 4 times a day. Telfa dressingswere applied, and soaked off, when removed.

The heparin solution relieved the pain. The deep irregular abrasions andthe areas where skin had been avulsed stopped weeping, revascularized,granulated, re-epithelialized and healed without contractures in 12days. The skin was comfortable and cosmetically pleasant, although someareas have no skin pigment.

EXAMPLE VII Healing of Abrasion-Avulsion of Palm Skin

A 40 year old male, injured in a motorcycle accident and suffering fromasphalt-pavement "burn" abrasions and avulsion of his right hand palmskin was treated with heparin. The initial topical dose of heparin was20,000 I.U.; then 5,000 I.U. or less, twice daily, for ten days, withheparin dose reduced or discontinued in revascularized areas wherebleeding could occur if heparin were continued.

The pain in the wound was relieved promptly by the topical applicationof the heparin. The wound was smaller, drier, and nonpainful one dayafter heparin was started. The wound revascularized, granulated,re-epithelialized and was healed 3 weeks post-accident. FIGS. 3A-D showthe palm, prior to heparin, after 1 and 4 days of heparin topicaltreatment, and healed on day 22.

EXAMPLE VIII Knee Skin Abrasion-Avulsion Cellulitis

A 20 year old male suffering from a knee injury was treated withheparin. Treatment consisted of parenteral and topical heparin, drynonadhering dressings and oral antibiotics. Subcutaneous heparin dosewas 20,000 I.U. daily, for 2 days, and 10,000 I.U., for 1 day. Topicalheparin dose was 20,000 I.U., initially, and 10,000 I.U., twice daily,reduced, progressively to 2,500 I.U., for 35 days. Antibiotics wereerythromycin, 500 mg, twice daily, for 2 days; then Keflex 500 mg, fourtimes daily, for 4 days; then Tegopen 500 mg, three times a day, for 10days; then erythromycin, 500 mg, twice daily, for 7 days; and finally,Ultracef 500 mg, twice daily, for 3 days.

Healing progressed slowly and unevenly through revascularization,granulation, and epithelialization of concentrically reducing circularareas. The cosmetic and functional results were good. FIGS. 4A-C showthe lesion, treatment days 1 and 41, and two years later.

EXAMPLE IX Necrotic Weeping Dermatitis of Lip

A 70 year old male, presenting a persisting necrotic weeping dermatitisof the lower lip, was treated with heparin. The lip lesion, which hadbeen resistant to all treatment by a dermatologist, was healed within 5days after topical heparin was started. Heparin was dripped onto thelip, four or more times a day; and 10,000 I.U. units, applied in divideddose, were used daily for 3 days. No dressings were used. There was noother treatment. Lip pain and itching subsided and stopped. The weepingfrom the white necrotic surface stopped. The lip revascularized andrapidly re-epithelialized, without contracture. When healed, the lip wascosmetically normal.

EXAMPLE X Nonhealing Bypass-Vein-Donor-Site in a Diabetic

An 81 year old diabetic with ischemic heart disease and chronic anginadisplaying a superficial saphenous vein donor-site wound of his leftlower extremity below the knee resulting from coronary artery bypasssurgery was treated with heparin. Previously, four months ofconventional treatment, the incision site medial to the tibia in themid-leg was open, weeping, ischemic and purulent. FIG. 5A.

Topical heparin solution, 10,000 I.U., initially, and 2,500 I.U.,applied twice daily, for 29 days, resulted in revascularization and scabclosure of the lesion. After 4 days of heparin the lesion was smaller,nonweeping, revascularizing and much less purulent. FIG. 5B. After 23days of topical heparin use, healing was unusually slow compared withother lesions of similar size treated with topical heparin. The patientshowered daily, thus exposing it to shower water daily. The patient wasadvised to keep the lesion dry, and thereafter the healing increased,with a scab covering the lesion on day 29 of heparin therapy.Erythromycin, then Augmentin, and dicloxacillin were given orally, 250mg, 4 times daily. FIGS. 5A-B show this early heparin healing phase.

Thirty-three days later, the very thick scab that was still adherent,and which became repeatedly wet when he resumed daily showering withoutcovering it, was removed. An incompletely healed, purulent base wasrevealed. FIG. 5D. Topical heparin was started again. Twice-dailyapplication of less than 2,500 I.U. daily accelerated and completed thehealing. FIGS. 5D show the lesion after 6 days of topical heparin.

In this case, heparin was used in both an early and in a late healingphase. In the early phase, heparin increasedrevascularization-granulation-re-epithelialization. In the late phase ofhealing, heparin increased re-epithelialization.

It will be understood by those skilled in the art that the aboveexamples of the instant invention are provided as representative of themany hithertofore unknown medically beneficial therapeutic uses ofheparin, or chemically related molecules, and should not be construed aslimiting the invention. It will be appreciated that numerous other suchapplications are anticipated and fall within the scope of the amendedclaims.

We claim:
 1. A method for facilitating transplanting organs, tissues orcells, comprising:contacting said organs, tissues or cells with a firstcomposition consisting essentially of compounds selected from the groupconsisting of heparin, heparinoids, and heparin sulfate in a firstphysiologically acceptable solution having an acidic pH and at aconcentration of about 1,500-5,000 units per milliliter, prior toremoval of said organs, tissues or cells from a donor patient;contacting said organs, tissues or cell removed from said donor patientwith a second composition consisting essentially of compounds selectedfrom the group consisting of heparin, heparinoids or heparin sulfate ina physiologically acceptable solution having an acidic pH and at aconcentration of about 1,500-40,000 units per milliliter; andtransplanting said organs, tissues or cells into a recipient patient,said recipient patient being treated with a third composition consistingessentially of heparin, heparinoids or heparin sulfate in aphysiologically acceptable solution in an amount effective to inhibitrejection of said organs, tissues or cells from said recipient patientand reducing the requirement for immunosuppressive drugs.
 2. A method asdescribed in claim 1 wherein said first and second solutions have a pHof about 5.5.
 3. A method as in claim 1 wherein said contacting of saidorgans, tissues or cells with said second composition comprisingprolonging the length of time of viability of said organs, tissues orcell between the time of harvest from said donor patient and the time ofimplantation into said recipient patient, as compared to the length oftime of viability of said organs, tissues or cells without saidcontacting.
 4. A method as in claim 1 wherein said compound is heparin.5. A method for preventing cellular destruction of biopsied material,comprising:removing said material from a patient; and contacting saidmaterial with a composition consisting essentially of compounds selectedfrom the group consisting of heparin, heparinoids and heparin sulfate ina physiologically compatible solution, said composition having an acidicpH and a heparin, heparinoid or heparin sulfate concentration of about1,500-20,000 units per milliliter.
 6. A method for extending the usablelifetime of bodily fluids, comprising:removing said bodily fluids from apatient's body; and contacting said bodily fluids with a compositionconsisting essentially of compounds selected from the group consistingof heparin, heparinoids or heparin sulfate in a physiologicallycompatible solution, wherein said heparin, heparinoids or heparinsulfate is present in said bodily fluids at a concentration of about1,500-5,000 units per milliliter.
 7. A method as described in claim 6wherein said bodily fluids are selected from the group consisting ofblood, plasma, and serum.
 8. A method for extending the usable lifetimeof organs, tissue or cells for transplant, comprising:removing saidorgans, tissues or cells from a donor's body; and contacting saidorgans, tissues or cells with a composition consisting essentially ofcompounds selected from the group consisting of heparin, heparinoids orheparin sulfate in a physiologically compatible solution having anacidic pH, wherein said heparin, heparinoids or heparin sulfate iscontacted with said organs, tissues or cells in a concentration of about5,000-20,000 units per milliliter.